pmR-mCherry

pmR-mCherry is a mammalian expression vector designed to constitutively express a microRNA of interest. Transfected cells can be identified by the coexpression of mCherry, a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). Coexpression of mCherry and your microRNA of interest allows easy monitoring and/or selection of microRNA-expressing cells by fluorescence microscopy or flow cytometry. The excitation and emission maxima of the native mCherry protein are 587 nm and 610 nm, respectively.

The pmR-mCherry multiple cloning site (MCS) is positioned in the 3’UTR, downstream of the mCherry coding sequence. Expression of mCherry and microRNA precursors cloned into the MCS is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the mCherry sequence. Both the fluorescent protein and the microRNA are expressed from a single mRNA transcript, which is cleaved by Drosha and Dicer to generate the mature microRNA.